final resuls after researchers tweaked it all up but before handed to students.

Esse art. não se aplica para o crime continuado.

このドキュメントでは、EDK II プラットフォーム記述ファイル（DSC）形式について説明します。EDK ビルドツールは、EDK II 互換性パッケージに含まれています。EDK II モジュールまたは EDK II ビルドツールを使用するには、EDK II DSC ファイルと FDF ファイルを使用する必要があります。

This article didn't really surprise me. We know Facebook was formed as a Tinder-style sexual rating system, so information of women being exploited by Facebook employees isn't unexpected from this company. This article brought up two specific examples of men using their power as Facebook employees to track the locations of women in real time, but also notes 52 total employees being fired for abusing their access to users' information.

This article just reinforces my worries about social media companies and my privacy. The article explains that Facebook accidentally stored millions of passwords for people in plain text that thousands of employees could access. Usually passwords are supposed to be stored in an encrypted way so people can't see what they are. If Facebook was dumb enough to let something like this slip through, I don't trust that they are taking care of all the information that they have collected on me.

This reading of "Private message" lets the reader (me) know what private messaging is which is in turn, a form of private communication. They go on by explaining the two different types of private messages. The first one being IRCs which are through apps like facebook or instagram. This means that these messages are not the main thing occurring on the app. Since posting is the main thing occurring. The second one is PMs which is through apps like WhatsApp and kik. This is where the messages are occurring through apps that are specifically made to direct message one another

[i12] shows how “no user notification” after a breach undercuts self-defense: without notice, people rarely rotate credentials or enable 2FA in time. To match the chapter’s shared-responsibility theme, please add a 72-hour post-breach action checklist (email/password rotation, stop reuse, enable 2FA, high-value account resets) plus a disclosure-timeline template that platforms should follow.

After reading this article, I think its very important to be aware about scams, and not to click on anything shady. Here, the article discusses about how hackers were even able to manipulate QR codes in order to steal data from victims. This can be done with other methods too, as recently, I have seen a lot of text messages about "lost packages", or "vehicle fines", so its important to also double check before clicking on external sources and websites.

This reminds me of how nearly everything on school laptops are usually monitored. I feel like people shouldn't use work/school technology for anything personal or private because it is definitely not private. The article stated that companies usually monitor employees for security reasons, especialy when they deal with sensitive materials.

This article talks about the data breaching that happened to Facebook sometime before 2019. The personal information of about 530 million users were stolen, however Facebook decided not to notify users about this. Something like this could put enormous amounts of people in danger. Our data within apps like Facebook include things like phone numbers, personal information, and even our connections. This data leaking could put people in risk to scammers, blackmail, etc. For example, there have been numerous cases were celebrities have had their data/personal photos to the public which caused major controversies. I think it was very immature for Facebook not to share this to users in order to not cause major backlash for the company.

We usually trustfully hand over our data to technology companies, but often overlook the fact that these companies are also composed of people. Since people are human, they always have some ill-intentioned thoughts. This requires more advanced institutions such as the government to supervise them in order to protect people's rights. When such incidents occur, it indicates that there is a significant lack of regulatory participation in this field, and this is an urgent problem that needs to be addressed now.

While privacy laws exist, like any other rules, they need to be reinforces. If not, there is no reason for people or entities to obey and follow them. As such, entities without solid governing such as some branches of the government could have the ability to do as they wish. If certain bad actors gained control over the ability to monitor people through the web, they would be able to monitor and find large amounts of information about its population and target certain demographics.

The CIDER technique is a 5-step analysis method to find out how your technology can (or can't) provide use to diverse users who go beyond what an "average" user needs or wants. CIDER stands for critique, imagine, design, expand, repeat. Critique the assumptions you may have about your design and the people who may use it. Imagine how one of your assumptions can lead to the exclusion of a type of user. Design possible changes so your design doesn't rely on your chosen assumption. Expand your knowledge by seeing and sharing more ideas with your team. Repeat the imagine and design steps for other assumptions.

Catch me if you can is a Stephan Spielberg movie about a man who makes fraudulent checks and cashes them in for money. More specifically he is often stealing people's identities or badges to help with providing information for these checks. Displaying a great example of the privacy of information of the internet.

The movie Catch Me if You Can is a fantastic movie and personally I think it's Leonardo DiCaprio's best movie along with the Wolf on Wall Street. In this movie, he is able to complete one of the largest bank fraud heists ever by becoming a 'pilot'. He does it by falsely becoming a pilot and then forging checks to his name. Something like this would never be able to happen in today's age, there's just too much security and restrictions when it comes to money and taxes. I think the best part of the whole movie is at the end when instead of being sentenced to prison for life, he is able to work with the FBI and help catch other criminals who did the same thing as him.

This is an article about how Facebook employee abusing their access of data to harass women. The author of the book An Ugly Truth: Inside Facebook’s Battle for Domination wrote about the crises that happened within the Facebook company over the past five years. It is very scary to me how employee abuses their power and put women in such dangerous position; and the woman won’t be able to know how her information gets leaked.

This article outlines the potential hazards and dangers of parents sharing photos and content of their children before they are old enough to consent. I believe this issue has only become more relevant since this article was published in 2020, with the rise of "family channels" and family-related content online. Parents are going beyond just sharing pictures; they are forcing their children into staged content, making them constantly perform the occurrences of their daily lives.

I’m not surprised that large companies like Facebook are mishandling their users’ private information. What’s really troubling is that even when such companies are exposed, or even sued, the actual victims rarely receive meaningful compensation. It feels deeply unfair.

Sometime before August 2019, Facebook's database was breached and the personal information of 530 million people was stolen and made public. I find it shocking that the company can decide not to notify their users that their information was stolen.

This article is documenting that Facebook stored passwords of users in a way that made them accessible to around 200,000 employees. I don't fear my information being "leaked" due to lack of assets that are desirable but that could just be a lack of understanding but this did concern me.

This source discusses the right to privacy and each person's right to privacy. It discusses the law and the United States rights that make sure that each citizen has the right to have their own personal and private information and lives.

I read this report from NPR titled "After Data Breach Exposes 530 Million, Facebook Says It Will Not Notify Users". I was really shocked. The data of 53 million people was leaked, and yet the company chose not to notify the users? This made me realize that big companies claim to "value privacy", but when problems occur, their first reaction is usually to protect themselves rather than protect the users. After reading it, I will be more cautious about the personal information on social media. After all, sometimes "the sense of security" is just an illusion.

communication is used everywhere to help yourself and others understand concepts

Fascinating

House keeping. It's a drag.

cool

While I in no way think these invasions of privacy are ethical or justified, I do think there's something to be said about taking accountability online. What I really mean by that is, I feel like some of these things are intuitive as consequences of using such a universal platform like the Internet. The work system example feels very obvious to me, and I would never send or say something not work appropriate on my work or school email. On the other hand, all private messages being completely accessible to the owners of social media is kind of ridiculous.

This section makes me think on the internet nowadays, there's absolutely no way to keep your information to yourself. People's information is in so many different companies, and users would not know how their information is being used either. Users has no control over their own privacy although it's something about themselves.

Making Custom Typewriter Rubber Stamps<br /> by Sam on Neo-Renaissance<br /> accessed on 2025-10-22T14:12:54

• Internal documents reviewed by The New York Times show Amazon plans to automate up to 75% of its operations in the coming years.
• The company expects automation to replace or eliminate over 500,000 U.S. jobs by 2033, primarily in warehouses and fulfillment centers.
• By 2027, automation could allow Amazon to avoid hiring around 160,000 new workers, saving about 30 cents per package shipped.
• This strategy is projected to save $12.6 billion in labor costs between 2025 and 2027.
• Amazon’s workforce tripled since 2018 to approximately 1.2 million U.S. employees, but automation is expected to stabilize or reduce future headcount despite rising sales.
• Executives presented to the board that automation could let the company double sales volume by 2033 without needing additional hires.
• Amazon’s Shreveport, Louisiana warehouse serves as the model for the future: it operates with 25% fewer workers and about 1,000 robots.
• A new facility in Virginia Beach and retrofitted older ones like Stone Mountain, Georgia, are following this design, which may shift employment toward more temporary and technical roles.
• The company is instructing staff to use softer language—such as “advanced technology” or “cobots” (collaborative robots)—instead of terms like “AI” or “robots,” to ease concerns about job loss.
• Amazon has begun planning community outreach initiatives (parades, local events) to offset the reputational risks of large-scale automation.
• The company has denied that the documents represent official policy, claiming they reflect the views of one internal group, and emphasized ongoing seasonal hiring (250,000 roles for holidays).
• Analysts suggest this plan could serve as a blueprint for other major employers, including Walmart and UPS, potentially reshaping U.S. blue‑collar job markets.
• The automation push continues a trajectory started with Amazon’s $775 million acquisition of Kiva Systems in 2012, which introduced mobile warehouse robots that revolutionized internal logistics.
• Recent innovations include robots like Blue Jay, Vulcan, and Proteus, aimed at performing tasks such as sorting, picking, and packaging with minimal human oversight.
• Long-term, Amazon may require fewer warehouse workers but more robot technicians and engineers, signaling a broader shift in labor type rather than total employment.

The Second Brain Scam<br /> by [[Corey Carvalho]]<br /> accessed on 2025-10-22T13:44:26

This shows how news traveled slowly. Colonists didn’t always know what was really happening in Europe.

The army was about to kill him for stealing. What does that tell us about British rules and power over soldiers?

Why was punishment so harsh?

The soldiers were guarding supplies in cold weather near Louisbourg, Nova Scotia. This shows how tough army life was in the winter.

Colonial soldiers often joined for short times. The army made them stay longer, which made some unhappy with British control.

Clough is talking about the French and Indian War, when the British and colonists fought the French and Native Americans for land in North America.

This heading should be an H2. I would change the hidden H2 to "Math Link Column" or something (if it actually needs to be filled out) and make this the H2. Also the font looks wrong to me?

There are a lot of experiments and studies that require other people. The idea of morals and ethnics is something that should be discussed and solidified before doing anything at all.

I feel like this idea is brought up in a lot of conversations but people don't actually know how big of a problem this is. We miss out on important perspectives that couldn't have been found otherwise.

Allowing more diverse and unique practices and theories into science allowed it to grow beyond its rigid ways. This opened the doors to a "higher ceiling" of scientific thought.

This is important to understand since science is a practice solely based on evidence being used to prove theories. If the character of the scientist since the evidence is more grounded and important.

When reading about science experiments and findings, I always think about the people behind it and how they must've felt being able to replicate scientific findings knowing they did it right. I know for sure that I am the kind of person to feel great about that.

This is important because it shows how science is a collective team effort, not an individual one. It requires cooperation between individuals to bring up the best results.

This is something I wish to change one day, as I want science to be something completely objective and not influenced by benefit or corruptiion.

I am excited to enter the scientific field specifically for this reason, working togethre with other people to view many different inputs and possiblities.

This is what I find most interesting about science, theres so many things that change and infinite possiblities.

This connects to not having a fixed mindset when doing research. During the research process it is important to test your original hypothesis because you never know what alternative solution you could come up with after the previous prediction.

It is especially important for science to be shared freely. This is because it will help people form networks with each other to solve complicated solutions during these difficult times of our society.

I can see how this thing is saying that if you want a dogs attention the stick or ball will get there attention and show how he is there for the human when they need him

This section explains password storage well, but it should explicitly separate encryption from hashing: sites should store passwords with a salted, slow hash (e.g., bcrypt and Argon2), not reversible encryption. Reversible schemes mean one leaked key exposes all passwords; slow hashing makes credential-stuffing economically painful. Minimal user practice: password manager + unique long passwords + TOTP or hardware-key 2FA.

Not only social platforms, but also the security of the online space has risen to a national level. In recent years, with the development of the Internet, more and more information has been digitized. Even some confidential data is stored in databases. Although these databases are usually very secure, they will be the first targets to be attacked once a war breaks out. For example, before the war between Russia and Ukraine, the Russian Cyber Security Department had already begun to attack some official websites of Ukraine. Therefore, today, cyber security in the network space has become particularly important.

I could just read the article but I'll do that later. but basically, I've always been confused on when hackers release the passwords of a bunch or users of a website or similar. Not so much how they do it (I still don't know how) but more so how they share that information. Like, do they just share the passwords without it's respected user? In that case it wouldn't be absolutely terrible since you still wouldn't know which password is for what account, but a smart hacker to maybe use a bot to try each of the 153 million passwords on one account (would still take ages, but at least you have a finite number of passwords to try). Or, do hackers put up all the password along with the users in a massive spreadsheet? That would make sense, you can just look up an account to hack and hack it easily. But do they share this on public platforms like Reddit? Do they share it directly with each other? Do they post it on some sort of evil dark web place? I'll find out I guess.

I wonder if there is any way a company is able to store secure information in a way such that even despite a breach in security, the information is still inaccessible to hackers. Would it be possible for the information to be stored instead on the users platform, and whenever the server needs to access it, it will instead request from the users computer? While this is less convenient and I assume it would require more processing time, it would be more secure as all information is not centralized.

I find it interesting that this article was released in 2021, but the incidents cited took place in 2015. Does this mean these incidents in which Facebook employees abused their power were not open to the public for 6 years?

I can see how this is true. Meta using selling information to ad agencies can easily lead to leaks and scams from other sources.

I find it shocking that the personal information that you willingly give to social media platforms can be so easily stolen and posted publicly on the internet. That is a huge breach of trust as people put their trust into these companies to save and protect their personal information.

I think the sentence "Although we are concerned about information privacy, we often share information with social media platforms and believe they will safely keep these details" is particularly true. We always say we want to protect privacy, but in reality, we still readily click "agree" and hand over our information without hesitation. Seeing this sentence made me reflect a bit - perhaps we have become too accustomed to convenience, and thus have overlooked the aspect of security.

This post is very interesting because it does not technically have any means for a lawsuit. If celebrities who could have gotten their information leaked, that could ruin their phones with the number of people who try to contact them after their information is leaked.

Treningi kierowania uwagą, mogą przynieść rezultaty dopiero wtedy kiedy dla danej osoby zaczyna to być proste, bezwysiłkowe, albo przynajmniej nie trdune. Tak jak tutaj w HRV, poprawiło się dopiero u osób, które trnowałydłużej.

L-> https://gemini.google.com/app/5c3775cc92786caf?hl=pl

Uwaga kierowana na ciało w momencie kiedy jest to dla osoby trudne, może tłumić HRV (obciążenie poznawcze).

L-> https://gemini.google.com/app/5c3775cc92786caf?hl=pl

Uwaga kierowana na ciało w momencie kiedy jest to dla osoby trudne, może tłumić HRV (obciążenie poznawcze).

L -> https://gemini.google.com/app/5c3775cc92786caf?hl=pl

I think I will share this with the others on my teaching team - They are veteran teachers but the way this text puts things plainly and sets out to clearly identify a guidance plan to turn to when challenging behavior presents itself is important.

I can't imagine that anyone in a early childcare setting would resort to any of these methods. That's frightening - the only reason rules are set in place is usually a response to a challenging situation. I can't even allow my mind to envision any of this happening - that's sad and angering. Force feeding children?

Agreed - getting angry in response to challenging behavior is never helpful.

I like these ways of responding - the child is not separated or singled out but provided additional support as most young children need when challenging behavior is presenting itself. We do most of these things in our classroom. Although there is one child who benefits from a hug and we ask him to verbally request that to respect him needs and only give when he requests.

Although personally I keep physical contact to a minimum when a child is upset because it can sometimes escalate the situation but these guidelines are good to know to safeguard our team.

We don't use our reading area as a quiet space enough - this could help as a place to divert children who are experiencing big feelings to reset and learn coping skills.

I like this suggestion - Roleplay expectations. Children love imaginative play and asking what if - playing this out allows children the space to use their imaginations as well as better understand expectations.

This is such a simple technique that is sometimes overlooked because some people assume certain expectations are common sense because of the way we were taught at home or the home environment we establish for our children at home.

Modeling expected behavior during transitions, I think would be very effective in redirecting some of the challenging behaviors we have been encountering related to transitions and cleaning learning centers.

This is true. The analysis of the current classroom schedule and environment has been helpful in brainstorming additional supports and solutions for some of the challenging behavior we have been seeing. Clear expectations that are posted and followed through on is important.

I agree - I first off see the difference in the responsiveness and bond when I am gone from the classroom for a day because of my schedule. It effects learning outcomes and trust in me as an educator. However, I have also seen the positive side of this - where building trust with one of my students has allowed him to build trust and confidence in his own voice around me. He is a bilingual learner and hasn't been verbally responsive to the lead teacher or paraprofessional. I can imagine if he hadn't built a relationship in which he felt safe he would still be nonverbal the entire day.

A child can be intellectually beyond ready for class and emotionally not ready at all to build relationships and receive the information being taught.

I have never read this article with my teaching group but can already tell this will help us with a plethora of useful information to help us in this teaching journey. At the pre-K age, self-control is really non-existent, especially if the basis for those skills aren't being enforced at home. It's a challenge at this age because they are VERY cute, and I'm just the teacher, so I can image the challenge of establishing boundaries and teaching self-control at home is a large one.

I totally agree - it's a very tall order. Teaching children self control in a classroom is very different from teaching children when you are babysitting or teaching your own children. I do believe that because I do not ye have children, the techniques I am learning to be effective will also change the way I parent. But this skill is very hard to teach in a positive way.

1833

1829

Relating back to this, I agree but also think most of your privacy gets taken away when you enter social media platforms. From what I have seen, people who post lifestyle content usually always get doxed by a random unemployed person who has too much time on their hands and slows every part of the video down until they find what they need. So I think we lose our privacy both to the media platforms and to the people online

This makes me really think about how even things we think are "private", like our privates messages, really aren't. These social media companies have accesses to pretty much all our activity online/on their apps. For example, when you allow apps like Instagram and Tiktok to access your photos and videos, they pretty much can see everything in your camera roll. This also makes me wonder if these social media companies can use this as black mail to important people like celebrities and politicians. People who influence our world.

This reminds me about a recent online controversy in my community where a very famous hijab brand's owner had a previous racist picture resurface online. Even though she had apologized for it multiple times the people of the black muslim community don't feel comfortable buying from her because of her previous actions. Which brings me to my point that digital footprint doesn't ever leave no matter how private it may seem.

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Lung Cancer and Mesothelioma Immunotherapy Guidelines Expert Panel.

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Lung Cancer and Mesothelioma CPG Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines. Last updated May 2025.

Addendum Summary

The SITC Lung Cancer and Mesothelioma CPG includes an addendum incorporating updates to the treatment landscape made by the SITC Lung Cancer and Mesothelioma CPG Expert Panel. A detailed description of the addendum updates can be found here: https://jitc.bmj.com/content/13/7/e003956add1

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines

Update October 2022

In response to recent data regarding increased risk for MACE in patients with RCC receiving ICI + VEGF TKI therapy, the ICI-related Adverse Events Guideline was updated in the following location: * General Expert Panel Recommendations

Reference: Prospective Cardiovascular Surveillance of Immune Checkpoint Inhibitor-Based Combination Therapy in Patients With Advanced Renal Cell Cancer: Data From the Phase III JAVELIN Renal 101 Trial

Update 8-7-2021

To add additional guidance on the management of patients with ICI-related hypophysitis and severe compressive symptoms, the ICI-related Adverse Events CPG was updated in the following location: * Endocrine Toxicity Expert Panel Recommendations

Check headline styles--these seem off.

Button: Visit the Office of the President

??

??

Period at the end.

Needs period at the end.

This should be an H3.

This should be an H2.

Social media companies are probably mainly incentivized by profit. Social media companies can get a lot of money by selling user data to advertisers. They might also analyze user data to tailor the algorithm to them and get users to stay on the platform as long as possible.

I think that after reading this chapter there really is no "privacy" on the internet. Elon musk for example accessed all of the direct messages and hackers can truly access whatever they what whenever they want, it doesn't feel very private to me.

Social media companies have a lot to gain from selling private data. They can sell it to as many as are willing to pay, money being the main incentive. Companies and corporations are willing to spend a lot of money to get to know data about their potential customer, if they know the majority of their potential customers are x, then they will respond by catering to x. Do a lot of people face similar problems? Luckily for them WebSiteName has just the solution!

I have a lot of fears around privacy on social media. I worry that social media companies are constantly listening and tracking what I say and do. I worry that they have a profile on me with everything about me. And I believe they are doing these things because I will be talking about something with my friends or parents, and then I will get an ad for it on instagram. I didn't plug anything into instagram to make them know that I was thinking about it yet they will somehow know. This is the kind of thing I don't like about social media.

At the moment there are still a good amount of legal protections that restrict companies from being too flagrant with your privacy but over the lat few decades that has become more and more of a market is companies actually selling people's information. I also think that people don't like the idea of a company having all their information so maybe another thing stopping them is the public sentiment toward private companies doing these massive privacy violations. But there is a lot of money on the other side of that fence and more and more and more companies are deciding that they want to take that leap and violate privacy.

I like that each of the center buttons open in a new window, but opening in a new window without flagging that is getting us dinged in Silktide. Do we need to add an "open in new window" icon next to the text on anything that will do that?

Two typos in the CEWIT alt text.

This should be an H2.

These should all be H2s.

This should be an H2

The image on this features Far Beyond banners. We will need an updated image.

This may need some space above it? Maybe 10px?

Last Reviewed 10/19/2024 (v2.4 Update)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Urothelial Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines.

v2.4 Update Summary

• The FDA granted accelerated approval of enfortumab vedotin in combination with pembrolizumab for the treatment of adult patients with locally advanced or metastatic urothelial cancer in December 2023 [Ref 3, 158].

• The FDA granted approval of nivolumab in combination with cisplatin and gemcitabine as first-line treatment of adult patients with unresectable or metastatic urothelial carcinoma in March 2024 [Ref 6].

• Data have been reported indicating similar responses to ICI therapy regardless of FGFR3 mutation status for patients with metastatic urothelial cancer [Ref 159].

• The FDA granted approval of nogapendekin alfa inbakicept with BCG for adult patients with BCG-unreseponsive NMIBC with carcinoma in situ with or without papillary tumors in April 2024 [Ref 160].

v2.3 Update Summary

• Atezoluzimab for the treatment of cisplatin- and platinum-ineligible patients with mUC was voluntary withdrawn by the manufacturer in November, 2022 [Ref 161].

• The FDA granted approval of nadofaragene firadenovec for the treatment of BCG-unresponsive NMIBC with CIS with or without papillary tumors in December 2022 [Ref 162].

• The FDA granted accelerated approval of enfortumab vedotin with pembrolizumab for the treatment of patients with locally advanced or metastatic urothelial carcinoma who are ineligible for cisplatin-containing chemotherapy in April 2023 [Ref 3, 158].

v2.2 Update Summary * The indication of pembrolizumab for the treatment of patients with mUC ineligible for platinum-containing chemotherapy or with disease progression on or after platinum-containing chemotherapy was revised in August 2021 to no longer specify PD-L1 status for eligibility [Ref 3].

v2.1 Update Summary * The FDA granted approval of nivolumab for the treatment of patients with urothelial cancer who are at high risk of recurrence after undergoing radical resection in August 2021 [Ref 6].

See the highlighted text for updated content and more detailed information.

Playwrighting

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Urothelial Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines.

Update 8-17-22

Based on advances in the field, including the following three FDA approvals and ongoing trials, the RCC CPG has been updated throughout the entire manuscript:

(1) Pembrolizumab for the adjuvant treatment of RCC: In November of 2021, the FDA approved pembrolizumab (anti-PD-1) for the adjuvant treatment of resected RCC at intermediate-high or high risk of recurrence following nephrectomy, or following nephrectomy and resection of metastatic lesions.

Reference: FDA approves pembrolizumab for adjuvant treatment of renal cell carcinoma FDA press release

(2) Pembrolizumab plus lenvatinib for the treatment of advanced RCC: In August of 2021, the FDA approved pembrolizumab in combination with lenvatinib (a VEGF receptor tyrosine kinase inhibitor; TKI) for the first-line treatment of patients with advanced RCC (aRCC).

Reference: FDA approves lenvatinib plus pembrolizumab for advanced renal cell carcinoma FDA press release

(3) Nivolumab plus cabozantinib for the treatment of advanced RCC: In January of 2021, the FDA approved nivolumab (anti-PD-1) in combination with cabozantinib (a VEGF receptor TKI) for the first-line treatment of patients with aRCC.

Reference: FDA approves nivolumab plus cabozantinib for advanced renal cell carcinoma FDA press release

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Prostate Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines.

Update August 2022

Based on the tissue agnostic approvals for pembrolizumab for the treatment of TMB-H and MSI-H/dMMR solid tumors and dostarlimab for the treatment of dMMR solid tumors, and their approved companion diagnostics, the Prostate Cancer CPG has been updated in the following location: - Introduction

References: Pembrolizumab (KEYTRUDA) TMB-H tissue-agnostic approval

Pembrolizumab (KEYTRUDA) MSI-H/dMMR tissue-agnostic approval

Dostarlimab (JEMPERLI) dMMR tissue-agnostic approval

Update 8-24-22

Based on new approvals of immunotherapy agents for the treatment of prostate cancer and new data that has been published since its original publication, the Prostate CPG was updated in the following locations: - Figure 1 - Prostate Cancer Treatment Algorithm

v1.1 Update

Since guideline publication, this phase II study assessing neoadjuvant cemiplimab reported a pCR rate of 51% and a major pathologic response rate of 13% in 70 patients with resectable stage II, III, or IV (M0) CSCC. Notably, an additional 9 patients were treated but did not undergo surgery. (This regimen was not FDA-approved at the time of update v1.1). [Ref 179]

v1.1 Update

In addition to the FDA-approved ICIs described in this section, since guideline publication, neoadjuvant anti-PD-1 therapy prior to curative-intent surgery demonstrated efficacy in a phase II study. (This regimen was not FDA-approved at the time of update v1.1.) [Ref 179]

v1.1 Update

In addition to the approved ICIs for MCC described in this section, since guideline publication retifanlimab was granted accelerated approval by the FDA in March 2023 for the treatment of metastatic or recurrent locally advanced MCC. Approval was based on the POD1UM-201 trial (NCT03599713). The primary outcome measures for approval were ORR and DOR (Table 2). Safety was assessed in 105 patients with MCC, where retifanlimab was determined to be safe and well-tolerated. Serious AEs occurred in 22% of patients, and the most common serious AEs were fatigue, arrhythmia, and pneumonitis. Permanent discontinuation of therapy due to AEs occurred in 11% of patients. [Ref 177, 178]

Additionally, although not FDA-approved at the time of update v1.1, two studies combining an anti-PD-(L)1 ICI with an anti-CTLA-4 ICI have reported efficacy in patients with recurrent/metastatic MCC. [Ref 180, 181]

Last reviewed 3/19/2024 (v1.1 Update)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication and made with the approval of the SITC NMSC CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

v1.1 Update Summary

• The FDA granted accelerated approval for retifanlimab (anti-PD-1 ICI) for the treatment of adult patients with metastatic or recurrent locally advanced MCC in March 2023. The NMSC CPG was updated in the following locations: Introduction, Merkel Cell Carcinoma, Recommended Immunotherapies for MCC, Figure 1 – FDA-Approved ICI agents for NMSC, Table 2 – NMSC Landmark Clinical Trial Data Leading to FDA Approvals for ICIs for MCC and Novel Strategies and Promising Future Directions. [Ref 177, 178]

• Data have been reported demonstrating efficacy with neoadjuvant anti-PD-1 ICI therapy prior to curative-intent surgery in patients with CSCC. Based on these practice-changing data, the NMSC CPG was updated in the following locations: Based on these practice-changing data, the NMSC CPG was updated in the following locations: Recommended Immunotherapies for CSCC, and Novel Strategies and Promising Future Directions for CSCC. [Ref 179]

• Data have been reported demonstrating efficacy combining an anti-PD-(L)1 ICI with an anti-CTLA-4 ICI for patients with advanced MCC. Based on these practice-changing data, the NMSC CPG was updated in the following locations: Merkel Cell Carcinoma, Recommended Immunotherapies for MCC, and Novel Strategies and Future Directions for CSCC. [Ref 180, 181]

See highlighted text for updated content and more detailed information.

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Multiple Myeloma Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines.

Update 2-1-2022

Based on the FDA approval of daratumumab in combination with carfilzomib and dexamethasone for adult patients with relapsed or refractory multiple myeloma who have received 1 to 3 prior lines of therapy on November 30, 2021, the Multiple Myeloma CPG has been updated in the following locations: * Monoclonal Antibodies Expert Panel Recommendations * Multiple Myeloma Key Monoclonal Antibody Therapies Trials

Reference: Daratumumab + hyaluronidase-fihj (Darzalex Faspro, Janssen Biotech, Inc.) and carfilzomib (Kyprolis, Amgen, Inc.) plus dexamethasone press release

Update 4-9-2021

Based on the approval of idecabtagene vicleucel for the treatment of adult patients with relapsed or refractory multiple myeloma after four or more prior lines of therapy, including an immunomodulatory agent, a proteasome inhibitor, and an anti-CD38 monoclonal antibody, the Multiple Myeloma CPG has been updated in the following locations: * Background * CAR T cell Therapies

Reference: Idecabtagene vicleucel (ABECMA) FDA press release

Based on the approval of isatuximab-irfc in combination with carfilzomib and dexamethasone, for the treatment of adult patients with relapsed or refractory multiple myeloma who have received one to three prior lines of therapy, the Multiple Myeloma CPG has been updated in the following locations: * Monoclonal Antibody Therapies * Multiple Myeloma Key Monoclonal Antibody Therapies Trials

Reference: Isatuximab-irfc (SARCLISA) FDA press release

Update 8-5-2020

Based on the accelerated approval of belantamab mafodotin-blmf for adult patients with relapsed or refractory multiple myeloma who have received at least 4 prior therapies, including an anti-CD38 monoclonal antibody, a proteasome inhibitor, and an immunomodulatory agent, the Multiple Myeloma CPG was updated in the following locations: * Antibody-Drug Conjugates * Background

Reference: Belantamab mafodotin-blmf (BLENREP) FDA press release

Last reviewed 02/10/2025 (v1.4 Update)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Lymphoma Cancer Immunotherapy Guideline Expert Panel. More information on the SITC Guidelines can be found at sitcancer.org/guidelines.

v1.4 Update Summary

• The FDA granted approval of denileukin diftitox-cxdl for the treatment of patients with stage 1 to 3 relapsed/refractory cutaneous TCL after at least 1 prior systemic therapy in August 2024 [Ref 242-243].

• The FDA granted accelerated approval of epcoritamab for adult patients with relapsed or refractory FL after two or more lines of systemic therapy in June 2024 [Ref 244, 245].

• The FDA granted approval of lisocabtagene maraleucel for adult patients with relapsed or refractory mantle cell lymphoma MCL who have received at least two prior lines of systemic therapy, including a Bruton tyrosine kinase inhibitor (BTKi) in May 2024 [Ref 246].

• The FDA granted accelerated approval of lisocabtagene maraleucel for adults with relapsed or refractory FL who have received two or more prior lines of systemic therapy in May 2024 [Ref 246].

• The FDA granted accelerated approval of lisocabtagene maraleucel for adult patients with relapsed or refractory CLL or SLL who have received at least 2 prior lines of therapy, including a BTKi and a B-cell lymphoma 2 (BCL-2) inhibitor in March 2024 [Ref 246].

• The FDA granted accelerated approval of zanubrutinib with obinutuzumab for relapsed or refractory FL after two or more lines of systemic therapy in March 2024 [Ref 247, 248].

• The FDA granted accelerated approval of glofitamab for relapsed or refractory diffuse large B-cell lymphoma DLBCL, not otherwise specified (NOS) or LBCL arising from FL, after two or more lines of systemic therapy in June 2023 [Ref 249].

• The FDA granted accelerated approval of epcoritamab for relapsed or refractory DLBCL NOS, including DLBCL arising from indolent lymphoma, and high-grade B-cell lymphoma (HGBL) after two or more lines of systemic therapy in May 2023 [Ref 244].

• The FDA granted approval of polatuzumab vedotin with a rituximab product, cyclophosphamide, doxorubicin, and prednisone (R-CHP) for adult patients who have previously untreated DLBCL, NOS or HGBL and who have an International Prognostic Index (IPI) score of 2 or greater in April 2023 [Ref 250, 251].

• The FDA granted accelerated approval of mosunetuzumab for adult patients with relapsed or refractory FL after two or more lines of systemic therapy in December 2022 [Ref 252].

• Data have been reported on the safety and efficacy of lenalidomide + obinutuzumab for relapsed or refractory FL and marginal zone lymphoma [Ref 253, 254].

• Data have been reported on the safety and efficacy of nivolumab + AVD for patients with Hodgkin lymphoma [Ref 255].

• Data have been reported on the safety and efficacy of nivolumab + chemotherapy and pembrolizumab + chemotherapy for patients with relapsed or refractory Hodgkin lymphoma [Ref 256–258].

• Data have been reported on the safety and efficacy of BV-nivolumab for the treatment of R/R PMBCL [Ref 259].

v1.3 Update Summary

• The FDA granted approval of lisocabtagene maraleucel for the treatment of adult patients with LBCL who have refractory disease to first-line chemoimmunotherapy or relapse within 12 months of first-line chemoimmunotherapy; or refractory disease to first-line chemoimmunotherapy or relapse after first-line chemoimmunotherapy and are not eligible for hematopoietic stem cell transplantation (HSCT) due to comorbidities or age in June 2022 [Ref 246].

• The FDA granted accelerated approval of tisagenlecleucel for the treatment of adult patients with relapsed or refractory FL following 2 or more lines of systemic therapy in May 2022 [Ref 113].

• The FDA granted approval of axicabtagene ciloleucel for adult patients with LBCL that is refractory to first-line chemoimmunotherapy or relapses within 12 months of first-line chemoimmunotherapy in April 2022 [Ref 111].

• The FDA granted accelerated approval of axicabtagene ciloleucel for adult patients with relapsed or refractory FL after two or more lines of systemic therapy in March 2021 [Ref 111].

v1.2 Update Summary

• The FDA granted accelerated approval of loncastuximab tesirine-lpyl for the treatment of adult patients with relapsed or refractory LBCL after two or more lines of systemic therapy, including DLBCL NOS, DLBCL arising from low grade lymphoma, and HGBL in April 2021 [Ref 260].**

v1.1 Update Summary

• The FDA granted approval of lisocabtagene maraleucel for the treatment of R/R large B-cell lymphoma after two or more lines of systemic therapy, including DLBCL not otherwise specified (including DLBCL arising from indolent lymphoma), high-grade B-cell lymphoma, PMBL, and FL grade 3B in February 2021. [Ref 246, 261]

Last reviewed 6/16/23 (v1.1(A) update supplement)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC HCC CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

A supplementary immunotherapy treatment algorithm for HCC was generated based on the Expert Panel recommendations. It can be found on the SITC website here.

Last reviewed 6/16/23 (v1.1(A) update supplement)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC HCC CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

SITC published an addendum to the guideline (Addendum 1, update v1.1(A)) based on the approval of tremelimumab in combination with durvalumab for adult patients with unresectable HCC. A supplementary immunotherapy treatment algorithm for HCC was generated based on the Expert Panel recommendations. It can be found on the SITC website here.

Last Reviewed 11/15/2024 (v1.1 Update)

The information on this page provides a detailed overview of updates to the guideline content based on changes in the field. Updates to the guideline outlined below were made with the approval of SITC's Head and Neck Squamous Cell Carcinoma (HNSCC) Guideline Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

Update v1.1 Summary * The FDA approved pembrolizumab for patients with recurrent or metastatic cutaneous squamous cell carcinoma in June 2020 [Ref 108, 169].

• The FDA approved toripalimab in combination with cisplatin and gemcitabine for first-line treatment of patients with metastatic or recurrent locally advanced nasopharyngeal carcinoma, or as a monotherapy for treatment of adult patients with recurrent, unresectable, or metastatic nasopharyngeal carcinoma with disease progression on or after platinum-containing chemotherapy in October 2023 [Ref 170, 171].

• Practice-changing data have been reported from CheckMate 141 regarding nivolumab as a first-line treatment in recurrent or metastatic HNSCC after progressing on platinum therapy for locally advanced disease in the adjuvant or primary (ie, with radiation) setting [Ref 172].

• Practice-changing data have been reported from KEYNOTE B10 and the FRAIL-IMMUNE/GORTEC 2018-03 trials, demonstrating efficacy of combining an anti-PD-(L)1 immune checkpoint inhibitor (ICI) with carboplatin + paclitaxel in frail patients with R/M HNSCC [Ref 173, 174].

Last Reviewed 5/25/24 (v1.1 Update)

The information on this page provides a detailed overview of updates to the guideline content based on changes in the field. Updates to the guideline outlined below were made with the approval of SITC's Gynecology Cancer CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

v1.1 Update Summary

• The FDA granted approval of pembrolizumab with chemoradiotherapy FIGO 2014 Stage III-IVA cervical cancer on January 12, 2024. Based on this approval, the Gynecologic Cancer CPG has been updated in the following location: Immunotherapy for the treatment of cervical cancer - Recommended immunotherapy treatments for cervical cancer (including Table 2). [Ref 13, 299, 300]

• The FDA granted approval of dostarlimab with carboplatin and paclitaxel, followed by single-agent dostarlimab for dMMR or MSI-H (as determined by an FDA-approved test) primary advanced or recurrent endometrial cancer on July 31, 2023. Based on this approval, the Gynecologic Cancer CPG has been updated in the following location: Immunotherapy for the treatment of endometrial cancer - Recommended immunotherapy treatments for endometrial cancer (including Table 3). [Ref 15, 121, 301]

See highlighted text for updated content and more detailed information.

Last reviewed 5/28/2025 (v1.1 Update)

Updates to the recommendations, tables, treatment algorithms, and/or guideline text in this publication are made with the approval of the SITC Gastrointestinal Cancer CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

v1.1 Update Summary

• The FDA approved nivolumab in combination with ipilimumab for adult and pediatric patients 12 years of age and older with unresectable or metastatic microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) colorectal cancer (CRC) in April 2025 [Ref 242, Ref 243].

• The FDA approved tislelizumab in combination with platinum and fluoropyrimidine-based chemotherapy for the first-line treatment of adult patients with unresectable or metastatic HER2-negative gastric or gastroesophageal junction adenocarcinoma whose tumors express PD-L1 (≥1) in December 2024 [Ref 229].

• The FDA approved zolbetuximab-clab in combination with platinum and fluoropyrimidine-containing chemotherapy for the first-line treatment of adults with locally advanced unresectable or metastatic HER2-negative gastric or gastroesophageal junction adenocarcinoma whose tumors are claudin (CLDN) 18.2 positive as determined by an FDA-approved test in October 2024 [Ref 230].

• The FDA approved tislelizumab for the treatment of adult patients with unresectable or metastatic esophageal squamous cell carcinoma (ESCC) after prior systemic chemotherapy that did not include a PD-L1 inhibitor in March 2024 [Ref 229].

• The indication of pembrolizumab with trastuzumab, fluoropyrimidine, and platinum-containing chemotherapy for the first-line treatment of patients with locally advanced unresectable or metastatic HER2-positive gastric or gastroesophageal junction (GEJ) adenocarcinoma was restricted to patients with CPS greater than or equal to 1, as determined by an FDA-approved test, in November 2023 [Ref 231].

• The FDA approved pembrolizumab with fluoropyrimidine- and platinum-containing chemotherapy as first-line treatment for adults with locally advanced unresectable or metastatic HER2-negative gastric or gastroesophageal junction (GEJ) adenocarcinoma in November 2023 [Ref 231].

• The FDA approved pembrolizumab with gemcitabine and cisplatin for the treatment of locally advanced unresectable or metastatic biliary tract cancer (BTC) in October 2023 [Ref 231].

• Additional changes made to address practice-changing data and updates in the field.

See the highlighted text for updated content and more detailed information.

v1.1 Update

Since guideline publication, additional neoadjuvant/pre-operative studies have shown efficacy with ICIs for patients with dMMR/MSI-H gastric/GEJ adenocarcinoma. These regimens were not FDA approved at the time of update v1.1 [Ref 96, 97, 160, 232, 233].

Last Reviewed 3/15/2024 (v1.2 Update)

The information on this page provides a detailed overview of updates to the guideline content based on changes in the field. Updates to the guideline outlined below were made with the approval of SITC's Breast Cancer CPG Expert Panel. More information on SITC Guidelines can be found at sitcancer.org/guidelines.

v1.2 Update Summary

• The FDA granted accelerated approval for dostarlimab for the treatment of dMMR recurrent or advanced solid tumors (along with the VENTANA MMR RxDx Panel companion diagnostic to detect dMMR) that have progressed on or following prior treatment and who have no satisfactory alternative treatment options on August 17, 2021. Based on these approvals, the Breast Cancer CPG has been updated in the following locations: Immunotherapy with PD-(L)1 Inhibitors for the Treatment of Advanced/Metastatic Breast Cancer and Diagnostics and Biomarker Testing in Patients with Advanced/Metastatic Breast Cancer. [Ref 290]

• The FoundationOne CDx assay was approved as the companion diagnostic for identifying patients with MSI-H tumors for treatment with pembrolizumab in February 2022. Based on this approval, the Breast Cancer CPG has been updated in the following locations: Diagnostics and Biomarker Testing in Patients with Advanced/Metastatic Breast Cancer. [Ref 291]

See highlighted text for updated content and more detailed information.

Note: This response was posted by the corresponding author to Review Commons. The content has not been altered except for formatting.

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Reply to the reviewers

Point-by-Point Response to Reviewers for Manuscript #RC-2024-02720

Manuscript Title: Molecular and Neural Circuit Mechanisms Underlying Sexual Experience-dependent Long-Term Memory in Drosophila.

Corresponding Author: Woo Jae Kim

We extend our sincere gratitude to the Managing Editor and both reviewers for their diligent and insightful evaluation of our manuscript. The comprehensive feedback provided has been invaluable, guiding us to significantly strengthen the manuscript's scientific rigor, logical cohesion, and overall impact. We have undertaken a substantial revision, incorporating new experimental evidence, reframing the central narrative, and improving data presentation to address all concerns raised.

The major revisions include:

1. New Experimental Evidence: We have performed three new sets of experiments to address key questions raised by the reviewers. First, we used the protein synthesis inhibitor cycloheximide to pharmacologically validate that the observed memory is indeed a form of long-term memory (LTM). Then, we performed genetic intersectional analyses to determine if the identified Yuelao (YL) neurons express the canonical sex-determination transcription factors doublesex (dsx) and fruitless (fru).
2. Narrative Reframing and Logical Restructuring: We fully agree with the reviewers that the logic of the original manuscript was confusing, particularly regarding the distinction between the broad Mushroom Body (MB) Kenyon Cell (KC) population and the specific YL neurons. The manuscript has been extensively rewritten to present a clear, hypothesis-driven narrative. We now frame the initial KC-related findings as part of a broader screening effort that logically led to the identification and focused investigation of the YL neuron circuit.
3. Refined Central Claim: Guided by the reviewers' feedback and our new data, we have sharpened our central claim. We now propose that YL neurons constitute a critical circuit for forming attractive taste- and pheromone-based memories derived from Gr5a neuronal inputs. This form of appetitive memory is distinct from the previously characterized internal reward state associated with ejaculation, adding a new layer to our understanding of how male flies remember and evaluate reproductive experiences.
4. Improved Data Quality and Analysis: In response to valid critiques, all imaging figures have been replaced with high-resolution versions. Furthermore, our methods for fluorescence quantification, particularly for the TRIC calcium imaging experiments, have been corrected to include normalization against an internal reference channel, adhering to established best practices. All requested genetic control experiments have been performed. We are confident that these comprehensive revisions have fully addressed all concerns and have transformed our manuscript into a much stronger, more focused, and logically sound contribution. We thank you again for the opportunity to improve our work and look forward to your evaluation of the revised manuscript.

Responses to Reviewer #1

General Comments: This study explores the molecular and neural circuitry mechanisms underlying sexual experience-dependent long-term memory (SELTM) in male Drosophila. The authors use behavioral, imaging, and bioinformatics approaches to identify YL neurons, a subset of mushroom body (MB) projecting neurons, as crucial for SELTM formation. They propose that YL neurons receive inputs from WG neurons via the sNPF-sNPFR pathway and implicate molecular players such as orb2, fmr1, MDAR2-CaMK, and synaptic plasticity in their function.

However, the evidence presented does not adequately support the authors' claims. The data fail to cohesively tell a logical story, and key conclusions appear to be based on assumptions and correlations rather than robust evidence.

• Answer: We are deeply grateful to both reviewers for their thorough and constructive evaluation of our manuscript. Their collective feedback has been instrumental in helping us to clarify the study's rationale, strengthen our interpretations, and significantly improve the overall quality and impact of the work. We appreciate the recognition of our study's potential to advance the understanding of how sexual experience modifies future mating behaviors and to elucidate the neuronal and molecular mechanisms of how memory regulates a key sexual behavior in male Drosophila*.

• *In response to the general comments, we have undertaken a major revision of the manuscript to improve the clarity, logic, and presentation. We have rewritten the Abstract and Introduction to more clearly define "sexual experience-dependent long-term memory" (SELTM) and articulate its significance in the context of adaptive decision-making and interval timing. The entire manuscript has been restructured to present a more logical, hypothesis-driven narrative that clearly distinguishes our initial broad screening from the focused investigation of the YL neuron circuit. We have also incorporated alternative interpretations of our data, particularly regarding the role of the YL circuit in regulating baseline mating duration in naive males, which has added more depth to the study. Finally, all figures have been remade in high resolution, and all requested genetic controls and methodological clarifications have been added to ensure rigor and reproducibility. We are confident that these revisions have addressed the reviewers' concerns and have resulted in a much stronger manuscript.

Comment 1: The study identifies the knowledge gap (lines 103-104) but fails to integrate relevant literature, particularly Shohat-Ophir et al., Science (2012), and Zer-Krispil et al., Curr Biol (2018). These studies established that ejaculation induces appetitive memory in male Drosophila via corazonin and NPF neurons. The current study does not provide direct evidence that the "act of mating itself" drives SELTM, as it includes both courtship and copulation.

Response: Thank you for highlighting these two landmark studies. We fully agree that Shohat-Ophir et al., Science (2012) and Zer-Krispil et al., Curr Biol (2018) were pivotal in demonstrating that ejaculation—and the accompanying corazonin/NPF signalling—can establish an appetitive memory in males.

In the revised manuscript we have now integrated both papers on lines 111-118:

“Previous work has shown that successful copulation is intrinsically rewarding to male Drosophila: a single mating encounter elevates brain neuropeptide F (NPF) levels and suppresses subsequent ethanol preference19. Importantly, Zer-Krispil et al. further demonstrated that ejaculation itself—artificially induced by optogenetic activation of corazonin (Crz) neurons—is sufficient to mimic this reward state, driving appetitive memory formation and up-regulation of NPF. These findings indicate that the act of ejaculation, rather than the entire courtship sequence, is the critical sensory event that gates post-mating reward.”

Comment 2: The nature of the observed long-lasting reduced mating duration requires clearer characterization: Is this an associative memory or experience-dependent behavioral plasticity? Can the formation of this long-term memory be blocked by protein synthesis inhibitors, such as cycloheximide?

Response: We thank the reviewer for this excellent suggestion to pharmacologically characterize the nature of the memory. To definitively test whether the observed SMD is a form of protein synthesis-dependent long-term memory (LTM), we performed a new experiment as suggested.

We have now included data in new Figure supplement 1I showing that feeding males the protein synthesis inhibitor cycloheximide (CXM) for 24 hours immediately following the sexual experience completely blocks the formation of the long-lasting SMD phenotype. Control flies fed a vehicle solution exhibited robust SMD. This result provides strong evidence that SELTM is not merely a form of transient behavioral plasticity but is a genuine form of LTM that requires de novo protein synthesis for its consolidation, a hallmark of LTM across species.[1]

The revised text was put on lines 173-176:

" To determine whether the persistent reduction in mating duration (SMD) depends on de-novo protein synthesis, we fed males the translational inhibitor cycloheximide (CXM). Under this regimen, CXM completely abolished the SMD phenotype (Fig. 1I)."

Comment 3: While schematics illustrate the working hypotheses, the text lacks detailed explanations, leaving the reader unclear about the rationale behind certain conclusions.

__Response: __Thank you very much for this insightful comment. We fully agree that the original manuscript did not provide sufficient textual justification for the conclusions derived from the schematics. In the revised version we have therefore added comprehensive explanations immediately following each figure (or schematic) that explicitly state the underlying rationale, the key observations supporting our hypotheses, and the logical steps leading to each conclusion. We believe these additions now make the reasoning transparent and easy to follow. We appreciate your feedback, which has substantially improved the clarity of our work.

• *

Comment 4*: The logic to draw certain conclusions was confusing and misleading. - For instance, the role of orb2 in SELTM is examined via knockdown in MB Kenyon cells (KCs) (using ok107>orb2-RNAi), which is irrelevant to the claim that orb2 functions in YL neurons. Additionally, RNAseq analyses (Fig. 1N-S) focusing on orb2 expression in a/b KCs are irrelevant to and cannot support the claim that Orb2 functions in YL neurons. *

*- Similarly, the claim (lines 302-303) that sNPF-R expression is exclusive to MB KCs conflicts with data showing effects when sNPF-R is knocked down in YL neurons. How can knocking-down a gene, which is exclusively expressed in neural population A, in neural population B affect a phenotype? This inconsistency undermines the interpretation of the results. *

*- Other examples include lines 223-227 and lines 246-249. It is very confusing how the authors came to the indications. *

- The authors also kept confusing the readers and themselves by mistakenly referring to MB KC a-lobe and YL a-lobe projection. They may know the difference between the two neural populations but they did not always refer to the right one in the text.

Response: We agree completely with the reviewer that the logic in the original manuscript was confusing and failed to clearly distinguish between the general MB Kenyon Cell (KC) population and the specific YL projection neurons. This was a major flaw, and we are grateful for the opportunity to correct it. We have undertaken a major revision of the manuscript's narrative and structure to present a clear, logical progression of discovery.

The new logical flow of the manuscript is as follows:

1. We first establish that sexual experience induces a robust, long-lasting SMD behavior that is dependent on protein synthesis
2. We then perform initial experiments to implicate the MB as a key brain region. We show that broad inhibition of MB KCs (using the ok107-GAL4 driver) disrupts SMD behavior.This result establishes the general involvement of the MB but lacks cellular specificity.
3. The remainder of the manuscript then focuses specifically on dissecting the molecular and cellular properties of these YL neurons. Finally, we have meticulously edited the entire manuscript to ensure that we always use precise terminology, clearly distinguishing between "YL neuron projections to the MB α-lobe" and the "MB KC α-lobe."

Comment 5*: The imaging figures provided are unfocused and poorly resolved, making it difficult to assess data quality. *

*- Colocalization analyses of orb2 and YL are unconvincing... Maximum intensity projection images are insufficient... complete image stacks with staining of orb2, YL, and KCs (MB-dsRed) are needed for validation. *

- Quantification of imaging data appears flawed. For example, claims of orb2 and CaMKII upregulation in MB a-lobe projections (e.g., Fig. S2F-J, Fig. 3M,N) are confounded by widespread increases in intensity across the brain, lacking specificity.

• *

*- The TRIC experiment analysis should normalize GFP signals to internal reference channel (RFP in the TRIC construct)... *

- In Fig. 6H-J, methods for counting synapse numbers are not described. How are synapse numbers counted in these low-resolution images?

Response: We sincerely apologize for the poor quality of the imaging data presented in the original manuscript. We agree with the reviewer's critiques and have taken comprehensive steps to rectify these issues.

• Image Quality: We apologize for not including the full image data in the original submission. The complete figure is now presented in revised Fig. 2J .
• Fluorescence Quantification: The fluorescence quantification has been re-analyzed. The Methods section now includes a detailed description of our protocol.
• TRIC Normalization: We apologize for not stating this explicitly in the previous version. As now described in the revised Methods subsection “Quantitative Analysis of Fluorescence Intensity”, all TRIC images were acquired with identical laser power and exposure settings. The GFP signal was background-corrected and then normalized to the RFP fluorescence encoded by the TRIC construct itself (UAS-mCD8RFP), which serves as an internal reference for construct expression and mounting thickness.

• Synapse Counting: We agree with the reviewer that the resolution of our images was insufficient for accurate synapse particle counting. We have therefore removed the problematic analysis from the former Fig 6H-J. Our conclusions regarding synaptic plasticity now rest on the more robust and quantifiable data showing a significant increase in the total area of dendritic (DenMark) and presynaptic (syt.eGFP) markers. Comment 6: The study presents data from unrelated learning paradigms (e.g., olfactory associative learning, courtship conditioning; Fig. 7) without justifying how these paradigms relate to SELTM. Particularly, the authors claimed that SELTM is related to Gr5a, which leads to appetitive memories, which involve PAM dopaminergic neurons and MB horizontal lobes. However, the olfactory associative learning with electric shock and courtship conditioning lead to aversive memories, that involve PPL1 dopaminergic neurons and the vertical lobes.

• *

Response: We thank the reviewer for requesting clarification on the rationale for including these experiments. The purpose of these assays was to test the specificity of the YL neuron circuit. A key question is whether YL neurons represent a general-purpose LTM circuit or one specialized for a particular memory modality.

The data show that knockdown of Orb2 or Nmdar2 specifically in YL neurons has no effect on the formation of LTM for aversive olfactory conditioning or aversive courtship conditioning. These negative results are critically important, as they demonstrate that the YL circuit is

not required for all forms of LTM. This finding strongly supports our revised central claim that YL neurons are specialized for processing appetitive memories derived from the specific sensory context of mating (i.e., taste and pheromonal cues from Gr5a neurons).

To improve the narrative flow of the main text, We rearranged the order of the articles. The relevant description is in lines 398-401:

“To determine whether YL neurons constitute a general LTM circuit or are dedicated to the appetitive context of mating, we tested two canonical aversive paradigms: electric-shock olfactory conditioning and courtship conditioning. If YL neurons serve as a universal LTM module, their genetic impairment should also impair aversive memory.”

lines 469-472:

“The inability of YL perturbation to impair aversive memories (Fig. 7) corroborates that this micro-circuit is dedicated to Gr5a-dependent SELTM rather than acting as a generic LTM hub”

Minor Issues

Comment 1: Fig 2F. YL projections are labeled as MBONs. Clarify whether YL neurons are the upstream or downstream (MBON) of KCs.

__Response: __Thank you for this helpful comment. As Huang et al., 2018[2] (Nat. Commun. 9:872) have mentioned, the MB093C-GAL4 driver is the MBON-α3 mushroom body output neuro. Consequently, YL neurons are positioned downstream of the MBON-α3.

We have now clarified this point in the revised manuscript lines 217-222:

“Each of these neurons extends a vertical fiber to the dorsal brain region, where they form dense arbors within the α-lobes of the mushroom body. Because the MB093C-GAL4 driver labels MBON-α3 output neuron[51], these YL arbors are positioned postsynaptically within the α-lobe and relay mushroom-body output to the anterior, middle, and posterior superior-medial protocerebrum.”

Comment 2: Extensive language polishing is required, as several sentences are unclear (e.g., lines 169-172).

Response: We apologize for the lack of clarity in the original text. The entire manuscript has undergone extensive revision and professional language editing to improve readability, precision, and grammatical accuracy.

Responses to Reviewer #2

Major Comments

Comment 1: Clearer articulation of the rationale, motivation, and significance of the overall study design and individual experiments can strengthen the manuscript and promote readership. For example, the beginnings of the abstract and introduction should define what authors mean by sexual experience-dependent long-term memory and its significance (including why it is "significant for reproductive success" (lines 46 and 92)). Similarly, employing more concrete language throughout the text will help anchor and contextualize the study. Interpretation is occasionally insufficient or does not follow directly from the data provided.

Response: We thank the reviewer for this valuable advice. We agree that the motivation and significance of our study were not articulated clearly enough. We have rewritten the Abstract and the beginning of the Introduction to address this. The revised text now explicitly defines SELTM as a protein synthesis-dependent, appetitive memory formed in response to gustatory and pheromonal cues. We explain its significance in the context of adaptive behavior, linking it to interval timing, a process by which male flies strategically adjust their mating investment (i.e., mating duration) based on prior experience to optimize reproductive success and energy expenditure. This framing provides a clearer context for our investigation into its underlying neural and molecular mechanisms.

Comment 2: Long term memory: I do not work on Drosophila memory, but a cursory search suggests that the field generally considers long term memory in Drosophila to last for 24 hr to days (courtship memory lasts for >24 hr). SMD decays between 12-24 hr after copulation. Could SMD be considered a short-term effect?

Response: This is an important point of clarification, as described in our response to Reviewer #1 (Major Comment 2), we have performed a new experiment demonstrating that the formation of SMD is blocked by the protein synthesis inhibitor cycloheximide (Figure 1I). This dependence on de novo protein synthesis is a defining characteristic of LTM, distinguishing it from short- and intermediate-term memory forms.[1] where memories lasting 12-24 hours are well-established as forms of LTM.[3] Therefore, based on both its duration and its molecular requirements, SMD represents a bona fide form of LTM.

The relevant statement is in lines 174-178:

"To determine whether the persistent reduction in mating duration (SMD) depends on de-novo protein synthesis, we fed males the translational inhibitor cycloheximide (CXM). Under this regimen, CXM completely abolished the SMD phenotype (Fig. 1I). This finding suggests that the reduction in mating investment is contingent upon the formation of LTM."

Comment 3: Fig 1B-E share the same control (naive) group. If these experiments were performed in the same replicate(s), they should be plotted in the same figure. If not, please provide more details on how experimental blocks were set up and how controls compared between replicates.

Response: Thank you for this helpful suggestion. We understand that sharing the same naive control across multiple panels (Fig. 1B–E) may raise concerns about data independence. However, we chose to present these panels separately for the following reasons:

1. Clarity and Readability: Each panel (B–E) represents a distinct temporal condition (0 h, 6 h, 12 h, 24 h post-isolation). Separating them avoids visual clutter and allows readers to focus on one time point at a time, improving interpretability.

__ Consistency with Internal Controls:__

Although the naive group is identical across panels, each experimental block (i.e., each isolation time point) was run independently on same days, with internal controls (naive vs. experienced) included in every block. This ensures that statistical comparisons remain valid within each panel, even if the naive data overlap.

We have now added a clear statement in the figure legend explaining that the naive group is shared across panels and that each time point was tested independently with internal controls. This maintains transparency while preserving the visual clarity of the current layout.

Comment 4: Serial mating (Fig 1F-H): please provide details on the methods. How much time elapsed between successive matings? Is a paired statistical test used? Sperm depletion also affects mating duration, and without this information the authors' conclusion (lines 155-156) does not automatically follow from the data.

Response:

1. __ Interval between successive matings__ We have rewritten the Methods to state explicitly that “as soon as one copulation ended the male was transferred immediately to a fresh virgin female, so the next mating began immediately.”

we add new method:

" Serial mating ____duration ____assay

Serial mating duration assay was identical to the standard procedure except that each male was presented with four DF virgin females in immediate succession: upon termination of the first copulation the male was immediately put into a fresh chamber containing the next virgin, the timer was restarted at first contact, and this step was repeated until four complete matings were recorded or 5 min elapsed without initiation, whichever came first."

__ Statistical test__

We apologize for omitting this detail. Unpaired t-test was used: for male the mating duration before (naïve) and after sexual experience was recorded, yielding paired observations. Prism’s unpaired t-test module was therefore applied to evaluate the mean difference.

The figure legend now states “with error bars representing SEM. Asterisks represent significant differences, as revealed by the Unpaired t test and ns represents non-significant difference (**p __ Mating duration versus sperm depletion__

We apologize for not having made it clear that these two observations are complementary, not contradictory. Previous work has shown that when male Drosophila copulate repeatedly, mating duration remains stable even though the number of sperm transferred—and thus the number of progeny sired—declines progressively [4]

The revised text is as follows (lines235-241):

"Previous work has shown that when male Drosophila copulate repeatedly, mating duration remains stable even though the number of sperm transferred—and thus the number of progeny sired—declines progressively. This dissociation confirms that the constant mating duration we observe in our serial-mating experiment (Fig. 1F–H) is consistent with normal sperm depletion and does not compromise the conclusion that the experience-dependent reduction in mating duration reflects long-term memory."

Thank you for helping us improve the clarity of our study.

Comment 5: Mating duration assay: Which isolation interval was chosen for the rest of the SMD experiments? The 12 hr en masse mating setup is relatively uncommon among studies on courtship/copulation/post-copulatory phenotypes, and introduces uncertainty and variability in the number and timing of matings that occurred during the 12 hr-window. This source of variability and its implication in interpreting the data should be acknowledged. Moreover, the 3 studies referenced in the methods all house males in groups of 4, whereas this study uses groups of 40. Could density confound the manifestation of SMD?

Response: We thank the reviewer for these important methodological questions.

• Isolation Interval: We have clarified in the Methods that virgin females were introduced into vials for last 1 day before assay.
• Housing Density: This is an excellent point. To control for any potential effects of housing density itself, we have clarified that our "naive" control males are also housed in groups of 40 for the same duration as the "experienced" males. Therefore, the only difference between the two groups is the presence of females, isolating the effect of sexual experience from the effect of social density. Comment 6: SMD behavior: comparing orb2 mutants and controls (Fig 1M and Fig S1K-L), loss of orb2 actually reduces the mating duration in native males (mean ~15 min) relative to controls (~20 min), and have possibly no effect on experienced males (~15 min). This is inconsistent with the SMD behavior demonstrated in Fig 1B-E. The same pattern is found for mushroom body silencing (Fig 1P, Fig S1M-N), orb2 knockdown in YL neurons (Fig 2D, Fig S2A-B), Fmr1 knockdown in YL neurons (Fig 3D, Fig S2B, S3D) and most other experiments where mating duration is not significantly different between naive and experienced males. This might demonstrate a separate role of YL neurons and its related circuit in regulating mating duration in naive males. Could the authors discuss this interpretation? As an aside, plotting genetic controls next to experimental groups is customary and facilitates comparisons between relevant groups.

Response: Thank you very much for this insightful observation.

1. Baseline differences among genotypes We agree that absolute mating duration differs slightly between genotypes (e.g. naive orb2∆/+ about 15 min vs. wild-type CS about 20 min). Such differences are common when mutations or transgenes are introduced into distinct genetic backgrounds, and they do not affect the within-genotype comparison that is the essence of SMD (sexual-experience-dependent shortening of mating duration). Therefore, for every experiment we compared naive vs. experienced males of the identical genotype, keeping all other variables constant.

Consistency of SMD across figures

In every manipulation that disrupts SMD memory (orb2∆, MB silencing, orb2-RNAi in YL neurons, Fmr1-RNAi in YL neurons, etc.) the naive–experienced difference disappears, whereas the genetic controls retain a significant ΔMD. This is fully consistent with Fig. 1B–E and demonstrates that the memory trace, not the basal duration, is abolished.

Figure layout

Following your suggestion, we have re-ordered all bar graphs so that the relevant genetic controls are placed immediately adjacent to the experimental groups, making within-panel comparisons easier.

We hope these clarifications and adjustments address your concerns.

Comment 7: Bitmap figures: unfortunately the bitmap figures are compressed and their resolution makes it difficult to evaluate the visual evidence.

Response: We apologize for the poor quality of the figures. All figures in the revised manuscript, including the scRNA-seq plots, have been remade as high-resolution vector graphics to ensure clarity and detail. For better understanding, different colored illustrations are also placed next to the scRNA-seq.

Comment 8: Sexual dimorphism of YL neurons: many neurons involved in sexual behaviors express dsx and/or fru. Do YL neurons express them?

Response: This is an excellent question. To address it, we performed a new set of experiments using genetic intersectional tools to test for the expression of doublesex (dsx) and fruitless (fru) in YL neurons. Our analysis, presented in figure supplement 2B, reveals that YL neurons are indeed fru-negative and dsx-negative. We therefore conclude that YL neurons do not belong to the canonical fru- or dsx-expressing neuronal classes and are unlikely to be intrinsically sex-specific.

We add explanation in lines 223-229:

"Our further analysis confirmed the presence of only three pairs of nuclei near the SOG in male brains, whereas female brains exhibit a greater number of nuclei near the AL (Fig. 2I), suggesting subtle sexual dimorphisms in GAL4MB093C-expressing neurons. Importantly, these neurons do not overlap with either fru- or dsx-expressing cells: co-immunostaining for GFP and Fru or Dsx revealed almost no colocalization in any brain region examined (Fig. S2B), indicating that YL neurons are distinct from the canonical sex-specific fru/dsx circuits."

Comment 9: Genetic controls for some crucial experiments are not provided, e.g. Fig 2J, Fig S3C, Fig S3E-F Fig 5B-C, F, Q-R, Fig S5A-E.

Response: We thank the reviewer for their careful attention to detail. We have now performed all the missing genetic control experiments.

Comment 10: Colocalization experiments: please provide more detail on how fluorescence is normalized for each channel across images, especially when the overall expression of an effector is up- or down-regulated after mating.

Response: We have updated the Methods section under "Quantitative Analysis of Fluorescence Intensity" and "Colocalization Analysis" to provide a detailed description of our normalization procedure.

Comment 11: Please resolve this apparent contradiction on the expression of Nmdar1 and 2 in YL neurons. On line 261: "both receptors co-expressing in Orb2-positive MB Kenyon cells"; on line 279-281 "Nmdar1 is not expressed with YL neurons [...] whereas Nmdar2 is expressed in a single pair of YL neurons..."

Response: We apologize for this contradiction, which arose from the confusing narrative structure of the original manuscript. As detailed in our response to Reviewer #1 (Major Comment 4), we have reframed the manuscript.

Comment 12: Particle analysis (Fig 6H-J): experienced males seem to have more synapses but trend towards smaller average size. It would be helpful to show number of synapses and average size as paired data, or show that the total particle area is larger in experienced males.

Response: We agree with the reviewer that this analysis was inconclusive and potentially misleading due to the limitations of image resolution. As noted in our response to Reviewer #1, we have removed this particle analysis (former Fig 6H-J) from the revised manuscript. Our claim for increased synaptic plasticity is now supported by the more robust measurement of the total fluorescence area of the pre- and postsynaptic markers, which shows a significant increase in experienced males.

Minor Comments

We thank the reviewer for their meticulous attention to detail. We have addressed all minor comments as follows:

Comment 1: 1. Some figures (e.g. Fig 3M-R) and experiments (e.g. oenocyte scRNA-seq) are not referenced in the text. dnc data is shown alongside amn and rut but the rationale for its inclusion is not provided.

__Response: __Original Fig. 3M-R (now Fig,3 M-O) was referenced on line 283. The rationale for including dnc data (as a canonical memory mutant) is now clarified in the text on lines 187-189:

"To ask whether the same molecular machinery underlies the SMD that follows sexual experience, we tested three classical memory mutants: dunce (dnc), amnesiac (amn), and rutabaga(rut)."

Comment 2: Some references might not point to the intended article (e.g. ref 123).

__Response: __The reference list has been checked and corrected.

Comment 3. Please plot genetic controls next to experimental genotypes as they are a crucial part of the experiment.

__Response: __All relevant figures now include plots of genetic controls next to experimental genotypes.

Comment 4. The "estimation statistics" plots are not necessary since the authors show individual data points. To further enhance data transparency, the authors may consider reducing the alpha and/or dot size so the individual data points are more readily visible.

Response: Thank you for this helpful suggestion! We fully agree that data transparency is essential. After carefully testing lower alpha values and smaller dot sizes, we found that either change markedly obscured the dense regions of the distributions. So we didn't change the size of the point.

The estimation-statistics overlays are kept for two courteous reasons: (i) they provide an immediate visual estimate of the mean difference and its 95 % confidence interval, which is the key statistic we base our conclusions on, and (ii) they spare readers from having to cross-reference separate tables.

Comment 5. For accessibility, please avoid using green and red in the same plot.

__Response: __We fully agree that red–green combinations can be problematic for colour-vision-impaired readers. In the present manuscript, however, the only panel that juxtaposes pure red and pure green is the Fly-SCOPE co-expression data. These scRNA-seq plots are provided only as supportive reference; the actual quantitative conclusions are based on independent genetic and imaging experiments that use magenta, cyan, yellow, and greyscale palettes. Moreover, the scope images are accompanied by detailed text descriptions of the overlapping cell clusters, so no essential information is lost even if the colours are indistinguishable

Comment 6. Fly Cell Atlas: please show color scales used for each gene as the color thresholds are gene-specific by default.The 3-color overlap on SCope also makes it very difficult to see the expression pattern for each gene. One possibility is outlining the Kenyon cells on the tSNE plots and showing the expression for each gene of interest.

Response: Thank you for this helpful suggestion. To avoid the ambiguity that arises from RGB blending in the three-colour overlay, we have added a small colour-mixing diagram next to the t-SNE plots (revised Fig. 1). This key shows the exact hues produced by pairwise and three-way overlaps:

• Red + Green = Yellow

• Red + Blue = Magenta

• Green + Blue = Cyan

• Red + Green + Blue = White

Thus, yellow, magenta or cyan dots indicate co-expression of two genes, while white dots mark cells where all three genes are detected. this diagram allows readers to interpret overlap colours at a glance without re-entering SCope.

Comment 7. Please also refer to Fly Cell Atlas as such. SCope is a visualization platform that houses multiple datasets.

__Response: __The reference to Fly Cell Atlas was added.

Comment 8. Please introduce acronyms and genetic reagents the first time they are mentioned.

__Response: __All acronyms and genetic reagents are now defined upon their first use.

Comment 9. Line 184: please specify "split-GAL4 reagents" instead of "advanced genetic tools".

__Response: __We have replaced "advanced genetic tools" with the more specific term "Split-GAL4 reagents."

Comment 10. Line 187: there are a few other lines with p>0.05 or p>0.01, so "uniquely" is inaccurate. Are the p-values in Table 1 corrected for multiple testing?

__Response: __The term "uniquely" has been revised for accuracy. No correction for multiple testing was applied because each entry in Table 1 represents a single pairwise comparison (naive vs. exp). Thus only one p-value was generated per experiment.

Comment 11. Some immunofluorescence panels lack scale bars.

__Response: __Scale bars have been added to all immunofluorescence panels.

Comment 12. Fig S2G-I: do authors mean "naive" instead of "group"?

__Response: __The term "group" in Fig S2G-I has been corrected to "naive."

Comment 13. Movie 1 should be referenced when YL neurons are first introduced.

__Response: __Movie 1 is now referenced when YL neurons are first introduced in the text.

Comment 14. Is Fig 4L similar to Fig 6L-N?

__Response: __This error has been corrected after the article was reformatted

Comment 15. Fig 7: please plot olfactory conditioning experiment results as either percentages, preference index, or paired numbers. "Number of flies/tube" is not as informative.

__Response: __Thank you for pointing this out. The bars in Fig. 7 indeed represent paired numbers, but we realise this was not stated explicitly. We apologize for the lack of clarity. In the revised manuscript we explained it in detail in figure legend and method. In the figure, we also marked the percentage of flies that chose to avoid the side of the stimulus with gas, and explained it in the Figure legend.

Reference

1. Lagasse F, Devaud J-M, Mery F. A Switch from Cycloheximide-Resistant Consolidated Memory to Cycloheximide-Sensitive Reconsolidation and Extinction in Drosophila. J Neurosci. 2009;29: 2225–2230. doi:10.1523/jneurosci.3789-08.2009
2. Huang C, Maxey JR, Sinha S, Savall J, Gong Y, Schnitzer MJ. Long-term optical brain imaging in live adult fruit flies. Nat Commun. 2018;9: 872. doi:10.1038/s41467-018-02873-1
3. Tonoki A, Davis RL. Aging Impairs Protein-Synthesis-Dependent Long-Term Memory in Drosophila. J Neurosci. 2015;35: 1173–1180. doi:10.1523/jneurosci.0978-14.2015
4. Macartney EL, Zeender V, Meena A, Nardo AND, Bonduriansky R, Lüpold S. Sperm depletion in relation to developmental nutrition and genotype in Drosophila melanogaster. Evol Int J Org Evol. 2021;75: 2830–2841. doi:10.1111/evo.14373

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Referee #2

Evidence, reproducibility and clarity

Summary:

Sun et al. show that Orb2-expressing, glutamatergic mushroom body neurons (YL neurons) are central to the "shorter mating duration (SMD)" behavior, where males reduce their mating duration up to 12 hours after the initial copulation. The authors use SMD as a model for understanding sexual experience-dependent long-term memory in males. A few genes implicated in long-term memory (Fmr1, CrebB) are required in YL neurons for SMD. The Nmdar-CaMKII signaling pathways is also implicated, and mating attenuates Ca2+ signaling and increases synaptic plasticity in the mushroom body and subesophageal ganglion.

Major comments:

1. Clearer articulation of the rationale, motivation, and significance of the overall study design and individual experiments can strengthen the manuscript and promote readership. For example, the beginnings of the abstract and introduction should define what authors mean by sexual experience-dependent long-term memory and its significance (including why it is "significant for reproductive success" (lines 46 and 92)). Similarly, employing more concrete language throughout the text will help anchor and contextualize the study. Interpretation is occasionally insufficient or does not follow directly from the data provided.
2. Long term memory: I do not work on Drosophila memory, but a cursory search suggests that the field generally considers long term memory in Drosophila to last for 24 hr to days (courtship memory lasts for >24 hr). SMD decays between 12-24 hr after copulation. Could SMD be considered a short-term effect?
3. Fig 1B-E share the same control (naive) group. If these experiments were performed in the same replicate(s), they should be plotted in the same figure. If not, please provide more details on how experimental blocks were set up and how controls compared between replicates.
4. Serial mating (Fig 1F-H): please provide details on the methods. How much time elapsed between successive matings? Is a paired statistical test used? Sperm depletion also affects mating duration, and without this information the authors' conclusion (lines 155-156) does not automatically follow from the data.
5. Mating duration assay: Which isolation interval was chosen for the rest of the SMD experiments? The 12 hr en masse mating setup is relatively uncommon among studies on courtship/copulation/post-copulatory phenotypes, and introduces uncertainty and variability in the number and timing of matings that occurred during the 12 hr-window. This source of variability and its implication in interpreting the data should be acknowledged. Moreover, the 3 studies referenced in the methods all house males in groups of 4, whereas this study uses groups of 40. Could density confound the manifestation of SMD?
6. SMD behavior: comparing orb2 mutants and controls (Fig 1M and Fig S1K-L), loss of orb2 actually reduces the mating duration in native males (mean ~15 min) relative to controls (~20 min), and have possibly no effect on experienced males (~15 min). This is inconsistent with the SMD behavior demonstrated in Fig 1B-E. The same pattern is found for mushroom body silencing (Fig 1P, Fig S1M-N), orb2 knockdown in YL neurons (Fig 2D, Fig S2A-B), Fmr1 knockdown in YL neurons (Fig 3D, Fig S2B, S3D) and most other experiments where mating duration is not significantly different between naive and experienced males. This might demonstrate a separate role of YL neurons and its related circuit in regulating mating duration in naive males. Could the authors discuss this interpretation? As an aside, plotting genetic controls next to experimental groups is customary and facilitates comparisons between relevant groups.
7. Bitmap figures: unfortunately the bitmap figures are compressed and their resolution makes it difficult to evaluate the visual evidence.
8. Sexual dimorphism of YL neurons: many neurons involved in sexual behaviors express dsx and/or fru. Do YL neurons express them? If they do, they might be a subset of characterized and named dsx/fru neurons.
9. Genetic controls for some crucial experiments are not provided, e.g. Fig 2J, Fig S3C, Fig S3E-F Fig 5B-C, F, Q-R, Fig S5A-E.
10. Colocalization experiments: please provide more detail on how fluorescence is normalized for each channel across images, especially when the overall expression of an effector is up- or down-regulated after mating.
11. Please resolve this apparent contradiction on the expression of Nmdar1 and 2 in YL neurons. On line 261: "both receptors co-expressing in Orb2-positive MB Kenyon cells"; on line 279-281 "Nmdar1 is not expressed with YL neurons [...] whereas Nmdar2 is expressed in a single pair of YL neurons in both male and female brains".
12. Particle analysis (Fig 6H-J): experienced males seem to have more synapses but trend towards smaller average size. It would be helpful to show number of synapses and average size as paired data, or show that the total particle area is larger in experienced males.

Minor comments:

1. Some figures (e.g. Fig 3M-R) and experiments (e.g. oenocyte scRNA-seq) are not referenced in the text. dnc data is shown alongside amn and rut but the rationale for its inclusion is not provided.
2. Some references might not point to the intended article (e.g. ref 123).
3. Please plot genetic controls next to experimental genotypes as they are a crucial part of the experiment.
4. The "estimation statistics" plots are not necessary since the authors show individual data points. To further enhance data transparency, the authors may consider reducing the alpha and/or dot size so the individual data points are more readily visible.
5. For accessibility, please avoid using green and red in the same plot.
6. Fly Cell Atlas: please show color scales used for each gene as the color thresholds are gene-specific by default.The 3-color overlap on SCope also makes it very difficult to see the expression pattern for each gene. One possibility is outlining the Kenyon cells on the tSNE plots and showing the expression for each gene of interest.
7. Please also refer to Fly Cell Atlas as such. SCope is a visualization platform that houses multiple datasets.
8. Please introduce acronyms and genetic reagents the first time they are mentioned.
9. Line 184: please specify "split-GAL4 reagents" instead of "advanced genetic tools".
10. Line 187: there are a few other lines with p>0.05 or p>0.01, so "uniquely" is inaccurate. Are the p-values in Table 1 corrected for multiple testing?
11. Some immunofluorescence panels lack scale bars.
12. Fig S2G-I: do authors mean "naive" instead of "group"?
13. Movie 1 should be referenced when YL neurons are first introduced.
14. Is Fig 4L similar to Fig 6L-N?
15. Fig 7: please plot olfactory conditioning experiment results as either percentages, preference index, or paired numbers. "Number of flies/tube" is not as informative.

Significance

The manuscript describes an extensive and comprehensive set of experiments aimed at elucidating the role of a subset of mushroom body neurons in mediating a male post-mating sexual behavior, which the authors use as a model for sexual experience-dependent long-term memory. Long-term post-mating responses in females have been well characterized in Drosophila and other insects, but post-mating long term memory in males are less well understood despite a few studies reporting their importance in mating success. How males adjust their mating duration based on internal and external cues can reveal insights about decision making and interval timer mechanisms. This study represents a functional advancement in the neuronal and molecular mechanisms of how memory and experience regulates a sexual behavior in male Drosophila. Overall, the manuscript can significantly benefit from general editing on clearer articulation of rationale and more appropriate interpretations of data. Higher resolution versions of bitmap figures is also crucial. The SMD experiments invite an alternative interpretation of data that centers on YL neurons' role on regulating mating duration in naive males, which alongside other roles of the mushroom body demonstrated in this manuscript, could add more depth to the study.

The findings in this manuscript will be of interest to a specialized audience interested in memory, neural circuits of behavior, and Drosophila sexual behavior. I work on Drosophila sexual behavior and circuits, but lacking experience on memory research, I am not as familiar with the mushroom body and conditioning experiments.

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Referee #1

Evidence, reproducibility and clarity

This study explores the molecular and neural circuitry mechanisms underlying sexual experience-dependent long-term memory (SELTM) in male Drosophila. The authors use behavioral, imaging, and bioinformatics approaches to identify YL neurons, a subset of mushroom body (MB) projecting neurons, as crucial for SELTM formation. They propose that YL neurons receive inputs from WG neurons via the sNPF-sNPFR pathway and implicate molecular players such as orb2, fmr1, MDAR2-CaMK, and synaptic plasticity in their function.

However, the evidence presented does not adequately support the authors' claims. The data fail to cohesively tell a logical story, and key conclusions appear to be based on assumptions and correlations rather than robust evidence.

Major comments:

1. The study identifies the knowledge gap (lines 103-104) but fails to integrate relevant literature, particularly Shohat-Ophir et al., Science (2012), and Zer-Krispil et al., Curr Biol (2018). These studies established that ejaculation induces appetitive memory in male Drosophila via corazonin and NPF neurons. The current study does not provide direct evidence that the "act of mating itself" drives SELTM, as it includes both courtship and copulation.
2. The nature of the observed long-lasting reduced mating duration requires clearer characterization: Is this an associative memory or experience-dependent behavioral plasticity? Can the formation of this long-term memory be blocked by protein synthesis inhibitors, such as cycloheximide?
3. While schematics illustrate the working hypotheses, the text lacks detailed explanations, leaving the reader unclear about the rationale behind certain conclusions.
4. The logic to draw certain conclusions was confusing and misleading.
   • For instance, the role of orb2 in SELTM is examined via knockdown in MB Kenyon cells (KCs) (using ok107>orb2-RNAi), which is irrelevant to the claim that orb2 functions in YL neurons. Additionally, RNAseq analyses (Fig. 1N-S) focusing on orb2 expression in a/b KCs are irrelevant to and cannot support the claim that Orb2 functions in YL neurons.
   • Similarly, the claim (lines 302-303) that sNPF-R expression is exclusive to MB KCs conflicts with data showing effects when sNPF-R is knocked down in YL neurons. How can knocking-down a gene, which is exclusively expressed in neural population A, in neural population B affect a phenotype? This inconsistency undermines the interpretation of the results.
   • Other examples include lines 223-227 and lines 246-249. It is very confusing how the authors came to the indications.
   • The authors also kept confusing the readers and themselves by mistakenly referring to MB KC a-lobe and YL a-lobe projection. They may know the difference between the two neural populations but they did not always refer to the right one in the text.
5. The imaging figures provided are unfocused and poorly resolved, making it difficult to assess data quality.
   • Colocalization analyses of orb2 and YL are unconvincing, especially given that orb2 is well-documented in literature as expressed in MB a-KCs and YL projection wrapping MB a-lobe. Maximum intensity projection images are insufficient for confirming colocalization; complete image stacks with staining of orb2, YL, and KCs (MB-dsRed) are needed for validation.
   • Quantification of imaging data appears flawed. For example, claims of orb2 and CaMKII upregulation in MB a-lobe projections (e.g., Fig. S2F-J, Fig. 3M,N) are confounded by widespread increases in intensity across the brain, lacking specificity.
   • The TRIC experiment analysis should normalize GFP signals to internal reference channel (RFP in the TRIC construct), as per established protocols in the original paper.
   • In Fig. 6H-J, methods for counting synapse numbers are not described. How are synapse numbers counted in these low-resolution images?
6. The study presents data from unrelated learning paradigms (e.g., olfactory associative learning, courtship conditioning; Fig. 7) without justifying how these paradigms relate to SELTM. Particularly, the authors claimed that SELTM is related to Gr5a, which leads to appetitive memories, which involve PAM dopaminergic neurons and MB horizontal lobes. However, the olfactory associative learning with electric shock and courtship conditioning lead to aversive memories, that involve PPL1 dopaminergic neurons and the vertical lobes.
7. Some figures are not referred to in the text. For example, Fig S1 K and L (also, what's the difference between these two figures?) and Fig 3M-R. What is MB-V3 in Fig 4J-K?

Minor issues

1. Fig 2F. YL projections are labeled as MBONs. Clarify whether YL neurons are the upstream or downstream (MBON) of KCs.
2. Extensive language polishing is required, as several sentences are unclear (e.g., lines 169-172).

Significance

This study potentially advances our understanding of how sexual experience modifies future mating behaviors. While previous work has shown that mating induces appetitive memory in males, the mechanisms linking this memory to future mating behavior remain poorly understood. This work could provide valuable insights into these mechanisms, pending appropriate revisions.

His supervisor thought their was nothing wrong

This passage emphasizes the way film responded to the wider cultural modernization of the early 1900s through technology, realism, and emotional involvement. The employment of camera movement and editing registered new conceptions of seeing and experiencing space and time, mirroring the break-throughs in science and psychology. Film therefore became a signifier of modernity, transforming visual entertainment into an influential art form expressive of human complexity and social transformation.

This part shows that the crisis wasn’t just social but emotional. Uncertainty caused peoples strife after the war. Even the universe seemed unstable. This new worldview influenced all types of art, suggesting that truth is known and knowledge is limited.

Can we turn this featured section off for launch and have the events calendar go straight across?

Mike to provide.

Do we want to add scholarships here? Allie to talk to Jacqueline

Move to first column?

Add Magazine here

Make "Administration"

Mike to update this section

Images of the students need to be updated.

Adita's CTA needs to say "Learn More About Adita"

"See All Events" button (I couldn't highlight it, sorry) is going to a broken link. Please update to go to https://calendar.stonybrook.edu/

Can these each link to the event pages?

Link these three buttons (Majors and Minors, Schedule a Tour, Apply Now)

Change and link to "Facts and Rankings"

Change to "Areas of Study" and link to Majors & Minors

Make Major plural (Majors)

Change to Deadline for Early Action is Nov. 11, link to https://www.stonybrook.edu/undergraduate-admissions/apply/first-year.php

Link to Admissions page.

Broken link.

As faculty who are considering what it means to bring OEP into our classrooms in this particular moment (fall of 2025) this note about perpetrating harm unintentionally resonates with our conversations. Specifically, how do we balance the risks that are changing and evolving for putting information into the public sphere (eg. depending on your context individuals can be targeted)? What does the evolution of how AI uses our information that's publicly available mean for the risks of doing OEPs in our classes?

How the Trump administration is dramatically reshaping education in America<br /> by [[John Yang]] of PBS at PBS News Weekend accessed on 2025-10-22T12:01:15

Interview with Jennifer Smith Richards of ProPublica

Anti-DEI, Anti-CRT bills

The magical source behind your favorite pastrami sandwiches and chili dogs<br /> by [[By Josh Heller]] for LAist<br /> accessed on 2025-10-22T11:56:41

uitproberen

This article talks about the right to privacy, and everyone in this country has the right to privacy on the internet. The internet should protect users' privacy. This right encourage more users to participate in any social media. I personally agree with this right, and I feel safe with the right to privacy.

It seems like there is an overlap where some people think social media is news media, and they believe everything they read on social media as the real news sometimes.

I think it’s interesting how the video points out that ADHD is found from kids before they are 12. I would think if symptoms start coming in later closer to puberty, it wouldn’t be different. I would even think it’s possible for younger kids to have mask traits such as procrastination due to the help of their parents so it might spark more obvious signs when they reach adulthood when there are less people helping them succeed.

http://johnsargentbarnard.com/Ribbon_Shutter/

Interesting UI for IndieWeb Challenges or events like 100 Days (https://indieweb.org/100_days).

This made me curious, are there any programs trying to fund science just for science's sake, or are all the funders profit based?

I have never thought about science as being inherently social. I think this idea is contradictory to the personality type and abilities most people associate with scientists.

This is very real. Society's image of a scientist is still very much limited to a white man. I think part of this problem is also that women and people in marginalized communities are more likely to be convinced younger that they are not good at science and math by those around them, limiting their abilities and motivation young.

I find this interesting. This system encourages advertising, and I see how that could easily lead to disingenuous work.

https://www.reddit.com/r/RetroFuturism/comments/1nreyfx/letters_by_radio_science_and_invention_artist/

“Letters by radio”, Science and Invention, (Artist Unknown) 1922<br />

https://www.reddit.com/r/typewriters/comments/1o8lzdm/trying_to_id_a_typewriter_from_a_video_game/

Folklore (2007, PlayStation 3)

https://www.instagram.com/nprfreshair/reel/DNVlf2tMstg/?hl=en

Terry Gross reading a book has rendered it useless for others to read.

Dog earing of top corner for interesting sections or questions she may have for the author.

Dog earing bottom corner as an indicator of remembering facts for the intro or for sentences she wants to quote.

Uses front of book for connecting themes and focus, so she won't forget it.

Introductions/prologues for quick overviews of what the book is about and why they wrote it.

https://www.instagram.com/reel/DNYh6b_uBwd/?hl=en

Terry Gross reads slowly to start and speeds up as she continues. She annotates and dog-ears as she reads and then makes notes and questions after she's done.

<br /> via https://www.facebook.com/misfitsmarket/posts/1196992682465751

I think it’s strange that disabled people aren’t able to design for themselves considering they have first hand experience with most of the issues. One of the first parts of designing is to empathize with a certain demographic so most disabled people would be able to understand the problems. I don’t get how you would need higher qualifications for someone designing a ramp as a disabled person rather than a non-disabled designer.

Sounds like pedagogy. I think i'm still not getting the difference between inclusive design and universal design for learning.

one, extendto manyby focusing on what’suniversally important toall humans.

Downright philosophical.

1. Increased access2. Reduced friction3. More emotional contextThe impact of inclusive design is morethan just the products that people use. It’salso a shift in our mindset, methods, andbehaviors.

Realm of psychology, spirituality, libera; arts/

studying the family life cycle will help with creating family interventions (Purpose of article???)

If we want to research extended family's impact on ecological systems, we need to start here by developing our understanding of families better.

Still not much research on the impact of extended family on adolescent mood :(

Purpose for the article (???)

This info can improve the help black people get in therapy

Broadening our research and definition of families could reduce some of the negative effects extended family may have regarding socioeconomic status

This is going in the conclusion, all 4 sources back this up.

the negative effects aren't tied to race, but socioeconomic status (which can be tied back to race in some ways but getting into that gets us off track of the research question)

This is the only negative effect I've seen for children/adolescents

The negative effects seem to be geared more towards the adults than the adolescents

Black and White families can't be compared the same way

Affirm the importance of extended family in single parent homes

The benefits of extended family are shown clearly through black families based off origin, values, beliefs, and varying familial structures that are distinct from the white "standard"

researchers turned their focus to tradition and culture, and less of the white "norm"

Important quote to include

The main problems of research on black families based on the nuclear definition was caused by assumptions that a) all black people will possess cultural attributes to the same degree, b) research on lower socioeconomic black families represented the entire black population (they generalized all of their research) and c) That white and black people had a cultural equivalence and that their behaviors could be compared the same way based off of a common standard

New Yorker Staff. 2007. “The Typing Life: How Writers Used to Write.” The New Yorker. https://www.newyorker.com/magazine/2007/04/09/the-typing-life (October 22, 2025).

typewriter plays a central role in the story

source? original quote?

via Darren Herschler-Henry's book

看了直想按怒

羽兔盒 Frabbitbox 唉... 2h Reply 陳咚咚 專業是一回事，班級經營閣是另外一回事 3h Reply Tsiā Uán Tsin 垃圾老師 4h Reply Karen Cheng 古早無學校 去學漢學？ 3h Reply Harvey Chen 有時候，學校真的是不得已的…… 3h Reply

Good example of basic Teams bot interaction

Interesting option for showcasing capabilities / FAQ, etc. without needing to manage a separate website.

https://www.facebook.com/groups/705152958470148/posts/1098284832490290/

https://www.reddit.com/r/typewriters/comments/1o8o98j/comment/njx2pgo/?context=1

Haul Out the Halloween (2025), Hallmark Movie

via Aaron Mitchell at https://www.facebook.com/groups/705152958470148/posts/1102482705403836/

Typewriter Typebar Holding Rack by grifter7 - Thingiverse<br /> by [[grifter7]] on [[Thingiverse.com]]<br /> accessed on 2025-10-22T10:26:22

Tool: Typebar Sorting Rack

This not only affects their health but also their education. If a student cannot afford to buy glasses they will likely continue to struggle seeing the board in school. Similarly if a student has a hard time hearing, they will face disadvantages which can affect their academic performance. It is unfair for intelligent students to fall behind because they can't afford the luxury of getting medical attention.

Many, like my family have come in the pursuit of the American Dream. While I do think it's possible, it is much easier said than done, especially for those who come from a low-income. Not everyone has access to resources that can help guide those new to this country. Because of that, many left to fend for themselves, not knowing what is necessary to set themselves for success. Yes, many can find employment, but employment does not equate to the American dream.

pbtpocket<br /> by [[Kaveh Shahbazian]]<br /> accessed on 2025-10-22T10:25:25